Capillary electrochromatography of peptides and proteins

Bandilla, Dirk; Skinner, Cameron D.

doi:10.1016/j.chroma.2004.05.075

Cited by 56 publications

(41 citation statements)

References 87 publications

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“…Recent progress in CEC of peptides and proteins is presented in three reviews in [11,156,157]. The last one [157] dealing with the nature of the forces controlling selectivity in the CEC of peptides, comes to a conclusion that both for the uncharged and charged peptides a synergistic interplay occurs in CEC systems between adsorptive/ partitioning events and electrokinetically driven motion.…”

Section: Electrochromatographymentioning

confidence: 98%

“…Their applications to separation of peptides have been broadened, and CE, EKC, and CEC have established as a recognized counterpart and/or complement to the up to recently, most frequently, used techniques for peptide separations, different modes of HPLC. In addition to the above-mentioned review [9], the summarized advances of CE and CEC of peptides in the recent period can be found in other review articles [10][11][12] and in the "concentrated form" also in the recent special issue of Electrophoresis devoted to CE and CEC of peptides [13].…”

Section: Introductionmentioning

confidence: 99%

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Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

2006

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The article gives a comprehensive review on the recent developments in the applications of high-performance capillary electromigration methods, zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography, to analysis, preparation, and physicochemical characterization of peptides. The article presents new approaches to the theoretical description and experimental verification of electromigration behavior of peptides, covers the methodological aspects of capillary electroseparations of peptides, such as rational selection of separation conditions, sample preparation, suppression of peptide adsorption, new developments in individual separation modes, and new designs of detection systems. Several types of applications of capillary electromigration methods to peptide analysis are presented: conventional qualitative and quantitative analysis, purity control, determination in biomatrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid and sequence analysis, and peptide mapping of proteins. Some examples of micropreparative peptide separations are given and capabilities of capillary electromigration techniques to provide important physicochemical characteristics of peptides are demonstrated.

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Section: Electrochromatographymentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

2006

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“…In recent years CEC has gained importance as a micro high-performance liquid separation technique for peptides and proteins [1][2][3][4][5]. CEC, which is often described as a hybrid of CZE and LC [6,7], has demonstrated higher separation efficiencies and flow permeabilities than capillary HPLC [8][9][10][11][12][13][14][15][16][17][18][19][20][21][22].…”

Section: Introductionmentioning

confidence: 99%

Electrochromatographic retention of peptides on strong cation‐exchange stationary phases

Nischang¹,

Höltzel²,

Tallarek³

2010

Electrophoresis

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We analyze the systematic and substantial electrical field-dependence of electrochromatographic retention for four counterionic peptides ([Met5]enkephalin, oxytocin, [Arg8]vasopressin, and luteinizing hormone releasing hormone (LHRH) ) on a strong cation-exchange (SCX) stationary phase. Our experiments show that retention behavior in the studied system depends on the charge-selectivity of the stationary phase particles, the applied voltage, and the peptides' net charge. Retention factors of twice positively charged peptides ([Arg8]vasopressin and LHRH at pH 2.7) decrease with increasing applied voltage, whereas lower charged peptides (oxytocin and [Met5]enkephalin at pH 2.7, [Arg8]vasopressin and LHRH at pH 7.0) show a concomitant increase in their retention factors. The observed behavior is explained on the basis of electrical fieldinduced concentration polarization (CP) that develops around the SCX particles of the packing. The intraparticle concentration of charged species (buffer ions, peptides) increases with increasing applied voltage due to diffusive backflux from the enriched CP zone associated with each SCX particle. For twice charged and on the SCX phase strongly retained peptides the local increase in mobile phase ionic strength reduces the electrostatic interactions with the stationary phase, which explains the decrease of retention factors with increasing applied voltage and CP intensity. Lower charged and weaker retained peptides experience a much stronger relative intraparticle enrichment than the twice-charged peptides, which results in a net increase of retention factors with increasing applied voltage. The CP-related contribution to electrochromatographic retention of peptides on the SCX stationary phase is modulated by the applied voltage, the mobile phase ionic strength, and the peptides' net charge and could be used for selectivity tuning in difficult separations.

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“…Capillary electrochromatography (CEC) has also become a powerful separation technique because it provides high column efficiency and selectivity by combining the advantages from both high-performance liquid chromatography (HPLC) and CE [35,36]. In CEC, the flow can be driven by both hydrodynamics and electroosmosis [37,38].…”

Section: Discussionmentioning

confidence: 99%

Photobleaching‐based flow measurement in a commercial capillary electrophoresis chip instrument

Wang¹,

Sas²,

Jiang³

et al. 2008

Electrophoresis

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For microfluidic analytical instruments, a facile, fast, and accurate instrument test is highly demanded. The test includes the quantitative verification of the relationship between pressure drop and flow velocity for the hydrodynamic pump, between the electric voltage and electroosmotic flow (EOF) for the high-voltage supply, and the chip quality. The key point for the test is the measurement of the flow velocity. However, most currently available velocimetries cannot be directly used without any instrumental modification or adding extra instruments. We applied a recently developed Laser Induced Fluorescence Photobleaching Anemometer (LIFPA) for the instrument test through measuring fluid flow velocity in a microfluidic instrument with optical measurement without any modification and extra instrument. We have successfully used the method to test Caliper HTS 250 System from Caliper Life Sciences (Hopkinton, MA) with its own light source and detector. The experimental result demonstrates that this single-point method of measuring flow velocity can be easily used for accurate test of a microfluidic instrument in less than 10 min at extremely low cost without any modification and extra instrument.

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Capillary electrochromatography of peptides and proteins

Cited by 56 publications

References 87 publications

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

Electrochromatographic retention of peptides on strong cation‐exchange stationary phases

Photobleaching‐based flow measurement in a commercial capillary electrophoresis chip instrument

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